IVIg-DCs promote expansion of NK cells and induce CD56^brightCD16⁻CCR7⁺CXCR3⁺ lymph node–type NK cells. (A) CTRL-DCs (top plots) or IVIg-DCs (bottom plots) were cultured with CFSE-labeled allogeneic T cells and NK cells at the ratio of DC/T/NK of 1:48:12, and after 5 days proliferation was determined. (Dotplots) The 2 top quadrants show proliferation of NK cells (CD56⁺CD3⁻); the 2 bottom quadrants show proliferation of the T cells (CD56⁻CD3⁺). (Histograms) Proliferation of all NK cells (left histogram), CD56^bright NK cells (middle histogram), and CD56^dim NK cells (right histogram). The plots are representative of 1 of 4 experiments. Percentages indicate proportions of NK cells that have undergone at least one division. (B) After the 5-day culture of NK cells with IVIg-DCs, NK cells were stimulated with PMA and ionomycin (2.5 μg/mL and 0.5 μg/mL), and CD107a expression on CD56⁺CD16⁺ and CD56⁺CD16⁻ subsets was determined. (C) After 5 days of culture with CTLR-DCs, IVIg-DCs, or HSA-DCs, percentages of CD56^bright NK cells, and percentages of NK cells expressing CD16, KIR receptors CD158a and CD158b, lymph node homing chemokine receptors CCR7 and CXCR3, and natural cytotoxicity receptors NKG2A and NKp30 were determined. NK cells cultured with IVIg-DCs up-regulated CCR7 and CXCR3, and down-regulated CD16, the KIR receptors, and NKp30 (N = 4; Student t test for paired data, *P < .05 compared with NK cells cocultured with CTRL-DCs). The percentages of cells are displayed within reference to the baseline percentage of expression of the markers on the NK cells before addition to culture. No expression of CXCR3 on NK cells was detected at this time point. Error bars represent SD.